Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus

Marion Ritzi, Kristina Tillack, Jeannine Gerhardt, Elisabeth Ott, Sibille Humme, Elisabeth Kremmer, Wolfgang Hammerschmidt, Aloys Schepers

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

The sequential binding of the origin recognition complex (ORC), Cdc6p and the minichromosome maintenance proteins (MCM2-7) mediates replication competence at eukaryotic origins of DNA replication. The latent origin of Epstein-Barr virus, oriP, is a viral origin known to recruit ORC. OriP also binds EBNA1, a virally encoded protein that lacks any activity predicted to be required for replication initiation. Here, we used chromatin immunoprecipitation and chromatin binding to compare the cell-cycle-dependent binding of pre-RC components and EBNA1 to oriP and to global cellular chromatin. Pre-replicative-complex components such as the Mcm2p-Mcm7p proteins and HsOrc1p are regulated in a cell-cycle-dependent fashion, whereas other HsOrc subunits and EBNA1 remain constantly bound. In addition, HsOrc1p becomes sensitive to the 26S proteasome after release from DNA during S phase. These results show that the complex protein-DNA dynamics at the viral oriP are synchronized with the cell division cycle. Chromatin-binding and chromatin-immunoprecipitation experiments on G0 arrested cells indicated that the ORC core complex (ORC25) and EBNA1 remain bound to chromatin and oriP. HsOrc6p and the MCM2-7 complex are released in resting cells. HsOrc1p is partly liberated from chromatin. Our data suggest that origins remain marked in resting cells by the ORC core complex to ensure a rapid and regulated re-entry into the cell cycle. These findings indicate that HsOrc is a dynamic complex and that its DNA binding activity is regulated differently in the various stages of the cell cycle.

Original languageEnglish (US)
Pages (from-to)3971-3984
Number of pages14
JournalJournal of Cell Science
Volume116
Issue number19
DOIs
StatePublished - Oct 1 2003
Externally publishedYes

Fingerprint

Origin Recognition Complex
Replication Origin
DNA Replication
Human Herpesvirus 4
Chromatin
Cell Cycle
Minichromosome Maintenance Proteins
DNA
Chromatin Immunoprecipitation
Proteins
S Phase
Mental Competency

Keywords

  • Cell cycle
  • EBNA1
  • ORC
  • oriP
  • Replication

ASJC Scopus subject areas

  • Cell Biology

Cite this

Ritzi, M., Tillack, K., Gerhardt, J., Ott, E., Humme, S., Kremmer, E., ... Schepers, A. (2003). Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus. Journal of Cell Science, 116(19), 3971-3984. https://doi.org/10.1242/jcs.00708

Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus. / Ritzi, Marion; Tillack, Kristina; Gerhardt, Jeannine; Ott, Elisabeth; Humme, Sibille; Kremmer, Elisabeth; Hammerschmidt, Wolfgang; Schepers, Aloys.

In: Journal of Cell Science, Vol. 116, No. 19, 01.10.2003, p. 3971-3984.

Research output: Contribution to journalArticle

Ritzi, M, Tillack, K, Gerhardt, J, Ott, E, Humme, S, Kremmer, E, Hammerschmidt, W & Schepers, A 2003, 'Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus', Journal of Cell Science, vol. 116, no. 19, pp. 3971-3984. https://doi.org/10.1242/jcs.00708
Ritzi M, Tillack K, Gerhardt J, Ott E, Humme S, Kremmer E et al. Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus. Journal of Cell Science. 2003 Oct 1;116(19):3971-3984. https://doi.org/10.1242/jcs.00708
Ritzi, Marion ; Tillack, Kristina ; Gerhardt, Jeannine ; Ott, Elisabeth ; Humme, Sibille ; Kremmer, Elisabeth ; Hammerschmidt, Wolfgang ; Schepers, Aloys. / Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus. In: Journal of Cell Science. 2003 ; Vol. 116, No. 19. pp. 3971-3984.
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